Assessment of cell viability of Bhk-21 cells at low and high concentrations of Tunicamycin after transfection of Bcl-2 ER localized proteins
Presentation Type
Poster
Major
Biology
Abstract
It is known that the B-cell lymphoma (Bcl-2) family of proteins has the ability to signal for apoptosis under stressful conditions to the cell, as well as to arrest apoptosis. The ability for Bcl-2 proteins to do this is due to their homology and their interaction with proteins such as the inositol 1,4,5-triphosphate (IP3) receptor, a calcium channel in the endoplasmic reticulum (ER). Blocking the IP3 receptor with 2- Aminoethoxydiphenyl borate (2-APB) has proven to be sufficient to block apoptosis at low levels of ER stress. In this study, Bcl-2 was used in an attempt to block apoptosis at low levels of ER stress. ER localized Bcl-2 (ER-Bcl-2-GFP) plasmid constructs were used to transfect Baby Hamster Kidney (BHK21) cells then exposed t plow and high levels of ER stress in an attempt to replicate the rescue 2- APB had on the temperature sensitive hamster firbroblasts (tsBN7). It was shown that the over expression of ER-Bcl-2 was able to sufficiently rescue tsBN7 cells from apoptosis under low levels of ER stress through temperature shifts. Also, it was proven that BHK21 cells were sufficiently rescued by 2- APB (50μM) under low levels (30nM) of Tunicamycin induced ER stress. Though, after transfections, and 36 hour exposure to low (20nM-30nM) and high (200nM) levels of Tunicamycin induced apoptosis, ER-Bcl-2 was not able to significantly rescue BHK21 cells. Albiet, there was a trend in some of the trials, but no replicates allow for significance
Faculty Mentor
Jay Brewster
Funding Source or Research Program
Summer Undergraduate Research in Biology
Location
Waves Cafeteria, Tyler Campus Center
Start Date
21-3-2014 2:00 PM
End Date
21-3-2014 3:00 PM
Assessment of cell viability of Bhk-21 cells at low and high concentrations of Tunicamycin after transfection of Bcl-2 ER localized proteins
Waves Cafeteria, Tyler Campus Center
It is known that the B-cell lymphoma (Bcl-2) family of proteins has the ability to signal for apoptosis under stressful conditions to the cell, as well as to arrest apoptosis. The ability for Bcl-2 proteins to do this is due to their homology and their interaction with proteins such as the inositol 1,4,5-triphosphate (IP3) receptor, a calcium channel in the endoplasmic reticulum (ER). Blocking the IP3 receptor with 2- Aminoethoxydiphenyl borate (2-APB) has proven to be sufficient to block apoptosis at low levels of ER stress. In this study, Bcl-2 was used in an attempt to block apoptosis at low levels of ER stress. ER localized Bcl-2 (ER-Bcl-2-GFP) plasmid constructs were used to transfect Baby Hamster Kidney (BHK21) cells then exposed t plow and high levels of ER stress in an attempt to replicate the rescue 2- APB had on the temperature sensitive hamster firbroblasts (tsBN7). It was shown that the over expression of ER-Bcl-2 was able to sufficiently rescue tsBN7 cells from apoptosis under low levels of ER stress through temperature shifts. Also, it was proven that BHK21 cells were sufficiently rescued by 2- APB (50μM) under low levels (30nM) of Tunicamycin induced ER stress. Though, after transfections, and 36 hour exposure to low (20nM-30nM) and high (200nM) levels of Tunicamycin induced apoptosis, ER-Bcl-2 was not able to significantly rescue BHK21 cells. Albiet, there was a trend in some of the trials, but no replicates allow for significance